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. Author manuscript; available in PMC: 2016 May 23.
Published in final edited form as: Mol Pharm. 2015 May 4;12(6):1992–2000. doi: 10.1021/mp500797n

Figure 1.

Figure 1

Engineered PFO binder reversibly inhibits the hemolytic activity of PFO. (a) Schematic of PFO neutralization. Binders to PFO were engineered on the Fn3 scaffold using yeast surface display techniques and selected for those that inhibit PFO function. The binder prevents pore formation when it is associated with PFO but allows normal activity following dissociation, allowing reversible inhibition. (b) Reduced hemolytic activity of PFO in the presence of PFO binders. Fn3s 1.1 and 1.2 were precomplexed with PFO at saturating concentrations (3 μM and 300 nM, respectively) and maintained as such after red blood cells were added. Hemoglobin release was measured after a 30 min incubation at 37 °C. (c) Clone 1.2 was displayed on the surface of yeast and tested for its ability to bind fluorescently labeled, soluble PFO mutants at 10 nM. The relative fluorescence units were normalized to that measured with PFO.